Pyrazolotriazines as kinase inhibitors

ABSTRACT

In its many embodiments, the present invention provides a novel class of pyrazolo[1,5-a]triazine compounds as inhibitors of kinases such as, for example, cyclin dependent kinases, methods of preparing such compounds, pharmaceutical compositions containing one or more such compounds, methods of preparing pharmaceutical formulations comprising one or more such compounds, and methods of treatment, prevention, inhibition, or amelioration of one or more diseases associated with the kinases using such compounds or pharmaceutical compositions.

This application is a divisional of patent application Ser. No.11/064,044 filed Feb. 23, 2005, now U.S. Pat. No. 7,038,045, whichclaims priority from U.S. Provisional application Ser. No. 60/547,685filed Feb. 25, 2004.

FIELD OF THE INVENTION

The present invention relates to pyrazolo[1,5-a]triazine compoundsuseful as protein kinase inhibitors (such as for example, the inhibitorsof the cyclin-dependent kinases, mitogen-activated protein kinase(MAPK/ERK), glycogen synthase kinase 3(GSK3beta), checkpoint kinase-1(“CHK-1”), checkpoint kinase-2 (“CHK-2”), Aurora kinases, (e.g., AuroraA, B, and C), protein kinase B (e.g., serine/threonine kinases such asAKT1, AKT2 and AKT3), and the like), pharmaceutical compositionscontaining the compounds, and methods of treatment using the compoundsand compositions to treat diseases such as, for example, cancer,inflammation, arthritis, viral diseases, neurodegenerative diseases suchas Alzheimer's disease, cardiovascular diseases, and fungal diseases.This claims priority from U.S. Provisional application, Ser. No.60/547,685 filed Feb. 25, 2004.

BACKGROUND OF THE INVENTION

Protein kinase inhibitors include kinases such as, for example, theinhibitors of the cyclin-dependent kinases (CDKs), mitogen activatedprotein kinase (MAPK/ERK), glycogen synthase kinase 3(GSK3beta),checkpoint kinase-1 (“CHK-1”), checkpoint kinase-2 (“CHK-2”), Aurorakinases, (e.g., Aurora A, B, and C), protein kinase B (e.g.,serine/threonine kinase such as AKT1, AKT2 and AKT3), and the like.Protein kinase inhibitors are described, for example, by M. Hale et alin WO02/22610 A1 and by Y. Mettey et al in J. Med. Chem., (2003) 46222–236. The cyclin-dependent kinases are serine/threonine proteinkinases, which are the driving force behind the cell cycle and cellproliferation. Individual CDK's, such as, CDK1, CDK2, CDK3, CDK4, CDK5,CDK6 and CDK7, CDK8 and the like, perform distinct roles in cell cycleprogression and can be classified as either G1, S, or G2M phase enzymes.Uncontrolled proliferation is a hallmark of cancer cells, andmisregulation of CDK function occurs with high frequency in manyimportant solid tumors. CDK2 and CDK4 are of particular interest becausetheir activities are frequently misregulated in a wide variety of humancancers. CDK2 activity is required for progression through G1 to the Sphase of the cell cycle, and CDK2 is one of the key components of the G1checkpoint. Checkpoints serve to maintain the proper sequence of cellcycle events and allow the cell to respond to insults or toproliferative signals, while the loss of proper checkpoint control incancer cells contributes to tumorgenesis. The CDK2 pathway influencestumorgenesis at the level of tumor suppressor function (e.g. p52, RB,and p27) and oncogene activation (cyclin E). Many reports havedemonstrated that both the coactivator, cyclin E, and the inhibitor,p27, of CDK2 are either over- or underexpressed, respectively, inbreast, colon, nonsmall cell lung, gastric, prostate, bladder,non-Hodgkin's lymphoma, ovarian, and other cancers. Their alteredexpression has been shown to correlate with increased CDK2 activitylevels and poor overall survival. This observation makes CDK2 and itsregulatory pathways compelling targets for the development years, anumber of adenosine 5′-triphosphate (ATP) competitive small organicmolecules as well as peptides have been reported in the literature asCDK inhibitors for the potential treatment of cancers. U.S. Pat. No.6,413,974, col. 1, line 23— col. 15, line 10 offers a good descriptionof the various CDKs and their relationship to various types of cancer.

CDK inhibitors are known. For example, flavopiridol (Formula I) is anonselective CDK inhibitor that is currently undergoing human clinicaltrials, A. M. Sanderowicz et al, J. Clin. Oncol. (1998) 16, 2986–2999.

Other known inhibitors of the CDKs include, for example, olomoucine (J.Vesely et al, Eur. J. Biochem., (1994) 224, 771–786) and roscovitine (I.Meijer et al, Eur. J. Biochem., (1997) 243, 527–536). U.S. Pat. No.6,107,305 describes certain pyrazolo[3,4-b]pyridine compounds as CDKinhibitors. An illustrative compound from the '305 patent has theFormula II:

K. S. Kim et al, J. Med. Chem. 45 (2002) 3905–3927 and WO 02/10162disclose certain aminothiazole compounds as CDK inhibitors.

Pyrazolopyrimidines are known. For Example, WO92/18504, WO02/50079,WO95/35298, WO02/40485, EP94304104.6, EP0628559 (equivalent to U.S. Pat.Nos. 5,602,136, 5,602,137 and 5,571,813), U.S. Pat. No. 6,383,790, Chem.Pharm. Bull., (1999) 47 928, J. Med. Chem., (1977) 20, 296, J. Med.Chem., (1976) 19 517 and Chem. Pharm. Bull., (1962) 10 620 disclosevarious pyrazolopyrimidines. Other publications of interest are: WO03/101993 (published Dec. 11, 2003), WO 03/091256 (published Nov. 6,2003), and DE 10223917 (published Dec. 11, 2003). Additionally, pendingU.S. patent applications, Ser. Nos. 10/654,546, 10/653,776, 10/654,168,10/654,163, 10/653,868 and 10/776,988 disclose variouspyrazolopyrimidines.

Pyrazolotriazines are known. Some publications disclosingpyrazolotriazines are: WO 99/67247 (published Dec. 29, 1999)DE 2900288A1, WO 02/096348 (published Dec. 5, 2002), and WO 02/50079 (publishedJun. 27, 2002).

There is a need for new compounds, formulations, treatments andtherapies to treat diseases and disorders associated with CDKs. It is,therefore, an object of this invention to provide compounds useful inthe treatment or prevention or amelioration of such diseases anddisorders.

SUMMARY OF THE INVENTION

In its many embodiments, the present invention provides a novel class ofpyrazolo[1,5-a]triazine compounds as inhibitors of kinases such as forexample, cyclin dependent kinases, methods of preparing such compounds,pharmaceutical compositions comprising one or more such compounds,methods of preparing pharmaceutical formulations comprising one or moresuch compounds, and methods of treatment, prevention, inhibition oramelioration of one or more diseases associated with the kinases, e.g.,CDKs, using such compounds or pharmaceutical compositions.

In one aspect, the present application discloses a compound, orpharmaceutically acceptable salts, solvates or esters of said compound,said compound having the general structure shown in Formula III:

wherein:

-   R¹ is selected from the group consisting of H, alkyl, aryl,    heteroaryl, heteroarylalkyl, arylalkyl, NR⁶R⁷, cycloalkyl and    cycloalkylalkyl, wherein each of said alkyl, aryl, heteroaryl,    heteroarylalkyl, cycloalkyl, cycloalkylalkyl and arylalkyl can be    unsubstituted or optionally independently substituted with one or    more moieties which can be the same or different, each being    independently selected from the group consisting of halo, alkyl,    aryl, heteroaryl, heterocyclyl, trifluoromethyl, OR⁶, NR⁶R⁷, SR⁶,    SO₂R⁶, CN, SO₂N(R⁶R⁷) and NO₂;-   R² is alkyl, cycloalkyl, alkenyl, alkynyl, trifluoromethyl, —OR⁷,    —SR⁷, hydroxyalkyl, haloalkyl, aryl, heteroaryl, halo, CN, formyl,    nitro, alkylcarbonyl, aralkylcarbonyl, heteroaralkylcarbonyl, or    -alkylene-N(R⁸R⁹) (where R⁸ and R⁹ independently represent H or    alkyl, or R⁸ and R⁹ taken together with the nitrogen in —N(R⁸R⁹)    form a five- to seven-membered heterocycle);-   R³ is —NR⁴R⁵,

H, alkyl, alkylthio, aralkylthio, alkylsulfinyl, or aralkylsulfinyl;

-   R⁴ is alkyl, cycloalkyl or heterocyclyl, wherein each of said alkyl,    cycloalkyl and heterocyclyl can be unsubstituted or optionally    independently substituted with 1–4 substituents which can be the    same or different, each substituent being independently selected    from the group consisting of halo, alkyl, hydroxymethyl,    hydroxyethyl, hydroxypropyl, trifluoromethyl, OR⁶, NR⁶R⁷, SR⁶,    SO₂R⁶, CN, SO₂N(R6R7)and NO₂;-   R⁵ is H, alkyl, aryl, heteroaryl, arylalkyl, cycloalkyl,    heterocyclyl, acyl or heteroarylalkyl;-   R⁶is H, alkyl or aryl;-   R⁷ is H or alkyl;-   R¹⁰ is halo, alkyl, hydroxyalkyl, trifluoromethyl, OR⁶, NR⁶R⁷, SR⁶,    SO₂R⁶, CN, SO₂N(R⁶R⁷) or NO₂; and-   n is 0 to 4, and when n is 2–4, the n moieties can be the same or    different, each being independently selected,    with the following provisos:

(i) that when R² is C₁–C₄ alkyl and R⁵ is H, then R⁴ is not a C₁–C₄alkyl;

(ii) that when R² is halo, CN, formyl, nitro, alkylcarbonyl,aralkylcarbonyl, heteroaralkylcarbonyl, or -alkylene-N(R⁸R⁹), then: (a)R³ is not H, alkylthio, aralkylthio, alkylsulfinyl, aralkylsulfinyl, or—NR⁴R⁵, and (b) n is not 0; and (iii) that when R² is alkyl, cycloalkyl,alkenyl or alkynyl, then R³ is not NH(methyl), N,N(dimethyl), NH(acetyl)or N(methyl)(acetyl).

The compounds of Formula III can be useful as protein kinase inhibitorsand can be useful in the treatment and prevention of proliferativediseases, for example, cancer, inflammation and arthritis. They may alsobe useful in the treatment of neurodegenerative diseases suchAlzheimer's disease, cardiovascular diseases, viral diseases and fungaldiseases.

DETAILED DESCRIPTION

In one embodiment, the present invention disclosespyrazolo[1,5-a]triazine compounds which are represented by structuralFormula III, or a pharmaceutically acceptable salt, solvate or esterthereof, wherein the various moieties are as described above.

In another embodiment, R¹ is selected from alkyl, aryl, heteroaryl,heteroarylalkyl, arylalkyl or NR⁶R⁷ wherein each of said alkyl, aryl,heteroaryl, heteroarylalkyl and arylalkyl can be unsubstituted oroptionally independently substituted with one or more substituents whichcan be the same or different, each substituent being independentlyselected from the group consisting of halo, alkyl, aryl, heteroaryl,trifluoromethyl, OR⁶, NR⁶R⁷, SR⁶, SO₂R⁶, CN, SO₂N(R⁶R⁷) and NO₂.

In another embodiment, R² is alkyl, cycloalkyl, alkynyl,trifluoromethyl, —OR⁷, or —SR⁷.

In another embodiment, R³ is NR⁴R⁵,

In another embodiment, R⁴ is alkyl, cycloalkyl or heterocyclyl, whereineach of said alkyl, cycloalkyl or heterocyclyl can be unsubstituted oroptionally independently substituted with one or more moieties which canbe the same or different, each moiety being independently selected fromthe group consisting of halo, alkyl, hydroxymethyl, hydroxyethyl,hydroxypropyl, trifluoromethyl, OR⁶, NR⁶R⁷, SR⁶, SO₂R⁶, CN, SO₂N(R⁶R⁷)and NO₂.

In another embodiment, R⁵ is H, alkyl, aryl, heteroaryl, arylalkyl orheteroarylalkyl.

In another embodiment, n is 1 to 2.

In another embodiment, R⁶ is H, alkyl or aryl.

In another embodiment, R⁷ is H or alkyl.

In another embodiment, R¹⁰ is halo, alkyl, hydroxymethyl, hydroxyethyl,hydroxypropyl, trifluoromethyl, OH, NR⁶R⁷, SR⁶, SO₂R⁶, CN or SO₂NR⁶R⁷.

In an additional embodiment, R¹ is selected from the group consisting ofphenyl, imidazolyl, imidazolyl-N-oxide, pyridyl, pyridyl-N-oxide,pyrazinyl, pyrazinyl-N-oxide, phenethyl, pyridone, —(CH₂)-pyridyl,—(CH₂)-pyridyl-N-oxide, —(CH₂)-pyrazinyl, —(CH₂)-pyrazinyl-N-oxide,—(CH₂)-pyridone, and —(CH₂)-imidazolyl-N-oxide, wherein each of saidphenyl, imidazolyl, pyridyl, pyridone, and pyrazinyl can beunsubstituted or optionally independently substituted with one or moremoieties which can be the same or different, each moiety beingindependently selected from the group consisting of halo, methyl, ethyl,trifluoromethyl, OH, alkoxy, NH₂, SH, SO₂CH₃, CN and SO₂NH(CH₂)₂CH₃.

In an additional embodiment, R² is selected from the group consisting ofmethyl, ethyl, cyclopropyl, cyclobutyl, cyclopentyl, ethenyl, —CF₃,hydroxy, methoxy, and ethoxy.

In an additional embodiment, n is 1.

In an additional embodiment, R³ is selected from the group consistingof:

-   -   (i) piperidyl substituted with a hydroxymethyl or hydroxyethyl;    -   (ii) pyrazinyl substituted with a hydroxymethyl or hydroxyethyl;    -   (iii) pyrrolidinyl substituted with a hydroxymethyl or        hydroxyethyl;    -   (iv) cyclohexyl substituted with a hydroxymethyl or        hydroxyethyl;    -   (v) cyclopentyl substituted with a hydroxymethyl or        hydroxyethyl);    -   (vi) —N(H)(piperidyl substituted with a hydroxymethyl or        hydroxyethyl);    -   (vii —N(H)(cyclohexyl substituted with a hydroxymethyl or        hydroxyethyl);    -   (viii) —N(H)(cyclopentyl substituted with a hydroxymethyl or        hydroxyethyl);    -   (ix) —N(H)(pyrrolidinyl substituted with a hydroxymethyl or        hydroxyethyl); and    -   (x) —N(H)[CH (hydroxymethyl)(isopropyl)].

In an additional embodiment, R⁴ is alkyl or cycloalkyl.

In an additional embodiment, R⁵ is H.

In an additional embodiment, R⁶ is H or alkyl.

In an additional embodiment, R⁷ is alkyl.

In an additional embodiment, R¹⁰ is hydroxymethyl or hydroxyethyl.

In a further embodiment, R¹ is selected from the group consisting ofimidazolyl, imidazolyl-N-oxide, pyridyl, pyridyl-N-oxide, pyrazinyl,pyrazinyl-N-oxide, phenethyl, pyridone, —(CH₂)-pyridyl,—(CH₂)-pyridyl-N-oxide, —(CH₂)-pyrazinyl, —(CH₂)-pyridone, and—(CH₂)-pyrazinyl-N-oxide;

-   -   R² is selected from the group consisting of methyl, ethyl and        cyclopropyl;    -   n is 1;    -   R³ is selected from the group consisting of:

-   (i) piperidyl substituted with a hydroxymethyl or hydroxyethyl;

-   (ii) pyrazinyl substituted with a hydroxymethyl or hydroxyethyl;

-   (iii) pyrrolidinyl substituted with a hydroxymethyl or hydroxyethyl;

-   (iv) cyclohexyl substituted with a hydroxymethyl or hydroxyethyl;

-   (v) cyclopentyl substituted with a hydroxymethyl or hydroxyethyl);

-   (vi) —N(H)(piperidyl substituted with a hydroxymethyl or    hydroxyethyl);

-   (vii —N(H)(cyclohexyl substituted with a hydroxymethyl or    hydroxyethyl);

-   (viii) —N(H)(cyclopentyl substituted with a hydroxymethyl or    hydroxyethyl);

-   (ix) —N(H)(pyrrolidinyl substituted with a hydroxymethyl or    hydroxyethyl); and

-   (x) —N(H)[CH(hydroxymethyl)(isopropyl)];    -   R⁴ is alkyl or cycloalkyl;    -   R⁵ is H;    -   R⁶ is H or alkyl; and    -   R⁷ is alkyl.

Yet another embodiment discloses the inventive compounds shown in Table1.

TABLE 1

As used above, and throughout this disclosure, the following terms,unless otherwise indicated, shall be understood to have the followingmeanings:

“Patient” includes both human and animals.

“Mammal” means humans and other mammalian animals.

“Alkyl” means an aliphatic hydrocarbon group which may be straight orbranched and comprising about 1 to about 20 carbon atoms in the chain.Preferred alkyl groups contain about 1 to about 12 carbon atoms in thechain. More preferred alkyl groups contain about 1 to about 6 carbonatoms in the chain. Branched means that one or more lower alkyl groupssuch as methyl, ethyl or propyl, are attached to a linear alkyl chain.“Lower alkyl” means a group having about 1 to about 6 carbon atoms inthe chain which may be straight or branched. The term “substitutedalkyl” means that the alkyl group may be substituted by one or moresubstituents which may be the same or different, each substituent beingindependently selected from the group consisting of halo, alkyl, aryl,cycloalkyl, cyano, hydroxy, alkoxy, alkylthio, amino, —NH(alkyl),—NH(cycloalkyl), —N(alkyl)₂, carboxy and —C(O)O-alkyl. Non-limitingexamples of suitable alkyl groups include methyl, ethyl, n-propyl,isopropyl and t-butyl.

“Alkynyl” means an aliphatic hydrocarbon group containing at least onecarbon-carbon triple bond and which may be straight or branched andcomprising about 2 to about 15 carbon atoms in the chain. Preferredalkynyl groups have about 2 to about 12 carbon atoms in the chain; andmore preferably about 2 to about 4 carbon atoms in the chain. Branchedmeans that one or more lower alkyl groups such as methyl, ethyl orpropyl, are attached to a linear alkynyl chain. “Lower alkynyl” meansabout 2 to about 6 carbon atoms in the chain which may be straight orbranched. Non-limiting examples of suitable alkynyl groups includeethynyl, propynyl, 2-butynyl and 3-methylbutynyl. The term “substitutedalkynyl” means that the alkynyl group may be substituted by one or moresubstituents which may be the same or different, each substituent beingindependently selected from the group consisting of alkyl, aryl andcycloalkyl.

“Aryl” means an aromatic monocyclic or multicyclic ring systemcomprising about 6 to about 14 carbon atoms, preferably about 6 to about10 carbon atoms. The aryl group can be optionally substituted with oneor more “ring system substituents” which may be the same or different,and are as defined herein. Non-limiting examples of suitable aryl groupsinclude phenyl and naphthyl.

“Heteroaryl” means an aromatic monocyclic or multicyclic ring systemcomprising about 5 to about 14 ring atoms, preferably about 5 to about10 ring atoms, in which one or more of the ring atoms is an elementother than carbon, for example nitrogen, oxygen or sulfur, alone or incombination. Preferred heteroaryls contain about 5 to about 6 ringatoms. The “heteroaryl” can be optionally substituted by one or more“ring system substituents” which may be the same or different, and areas defined herein. The prefix aza, oxa or thia before the heteroarylroot name means that at least a nitrogen, oxygen or sulfur atomrespectively, is present as a ring atom. A nitrogen atom of a heteroarylcan be optionally oxidized to the corresponding N-oxide. Non-limitingexamples of suitable heteroaryls include pyridyl, pyrazinyl, furanyl,thienyl, pyrimidinyl, pyridone (including N-substituted pyridones),isoxazolyl, isothiazolyl, oxazolyl, thiazolyl, pyrazolyl, furazanyl,pyrrolyl, pyrazolyl, triazolyl, 1,2,4-thiadiazolyl, pyrazinyl,pyridazinyl, quinoxalinyl, phthalazinyl, oxindolyl,imidazo[1,2-a]pyridinyl, imidazo[2,1-b]thiazolyl, benzofurazanyl,indolyl, azaindolyl, benzimidazolyl, benzothienyl, quinolinyl,imidazolyl, thienopyridyl, quinazolinyl, thienopyrimidyl,pyrrolopyridyl, imidazopyridyl, isoquinolinyl, benzoazaindolyl,1,2,4-triazinyl, benzothiazolyl and the like. The term “heteroaryl” alsorefers to partially saturated heteroaryl moieties such as, for example,tetrahydroisoquinolyl, tetrahydroquinolyl and the like.

“Aralkyl” or “arylalkyl” means an aryl-alkyl-group in which the aryl andalkyl are as previously described. Preferred aralkyls comprise a loweralkyl group. Non-limiting examples of suitable aralkyl groups includebenzyl, 2-phenethyl and naphthalenylmethyl. The bond to the parentmoiety is through the alkyl.

“Alkylaryl” means an alkyl-aryl-group in which the alkyl and aryl are aspreviously described. Preferred alkylaryls comprise a lower alkyl group.Non-limiting example of a suitable alkylaryl group is tolyl. The bond tothe parent moiety is through the aryl.

“Cycloalkyl” means a non-aromatic mono- or multicyclic ring systemcomprising about 3 to about 10 carbon atoms, preferably about 5 to about10 carbon atoms. Preferred cycloalkyl rings contain about 5 to about 7ring atoms. The cycloalkyl can be optionally substituted with one ormore “ring system substituents” which may be the same or different, andare as defined above. Non-limiting examples of suitable monocycliccycloalkyls include cyclopropyl, cyclopentyl, cyclohexyl, cycloheptyland the like. Non-limiting examples of suitable multicyclic cycloalkylsinclude 1-decalinyl, norbornyl, adamantyl and the like, as well aspartially saturated species such as, for example, indanyl,tetrahydronaphthyl and the like.

“Halogen” means fluorine, chlorine, bromine, or iodine. Preferred arefluorine, chlorine and bromine.

“Ring system substituent” means a substituent attached to an aromatic ornon-aromatic ring system which, for example, replaces an availablehydrogen on the ring system. Ring system substituents may be the same ordifferent, each being independently selected from the group consistingof alkyl, alkenyl, alkynyl, aryl, heteroaryl, aralkyl, alkylaryl,heteroaralkyl, heteroarylalkenyl, heteroarylalkynyl, alkylheteroaryl,hydroxy, hydroxyalkyl, alkoxy, aryloxy, aralkoxy, acyl, aroyl, halo,nitro, cyano, carboxy, alkoxycarbonyl, aryloxycarbonyl,aralkoxycarbonyl, alkylsulfonyl, arylsulfonyl, heteroarylsulfonyl,alkylthio, arylthio, heteroarylthio, aralkylthio, heteroaralkylthio,cycloalkyl, heterocyclyl, —C(═N—CN)—NH₂, —C(═NH)—NH₂, —C(═NH)—NH(alkyl),Y₁Y₂N—, Y₁Y₂N-alkyl-, Y₁Y₂NC(O)—, Y₁Y₂NSO₂— and —SO₂NY₁Y₂, wherein Y₁and Y₂ can be the same or different and are independently selected fromthe group consisting of hydrogen, alkyl, aryl, cycloalkyl, and aralkyl.“Ring system substituent” may also mean a single moiety whichsimultaneously replaces two available hydrogens on two adjacent carbonatoms (one H on each carbon) on a ring system. Examples of such moietyare methylene dioxy, ethylenedioxy, —C(CH₃)₂— and the like which formmoieties such as, for example:

“Heterocyclyl” means a non-aromatic saturated monocyclic or multicyclicring system comprising about 3 to about 10 ring atoms, preferably about5 to about 10 ring atoms, in which one or more of the atoms in the ringsystem is an element other than carbon, for example nitrogen, oxygen orsulfur, alone or in combination. There are no adjacent oxygen and/orsulfur atoms present in the ring system. Preferred heterocyclyls containabout 5 to about 6 ring atoms. The prefix aza, oxa or thia before theheterocyclyl root name means that at least a nitrogen, oxygen or sulfuratom respectively is present as a ring atom. Any —NH in a heterocyclylring may exist protected such as, for example, as an —N(Boc), —N(CBz),—N(Tos) group and the like; such protections are also considered part ofthis invention. The heterocyclyl can be optionally substituted by one ormore “ring system substituents” which may be the same or different, andare as defined herein. The nitrogen or sulfur atom of the heterocyclylcan be optionally oxidized to the corresponding N-oxide, S-oxide orS,S-dioxide. Non-limiting examples of suitable monocyclic heterocyclylrings include piperidyl, pyrrolidinyl, piperazinyl, morpholinyl,thiomorpholinyl, thiazolidinyl, 1,4-dioxanyl, tetrahydrofuranyl,tetrahydrothiophenyl, lactam, lactone, and the like.

It should be noted that in hetero-atom containing ring systems of thisinvention, there are no hydroxyl groups on carbon atoms adjacent to a N,O or S, as well as there are no N or S groups on carbon adjacent toanother heteroatom. Thus, for example, in the ring:

there is no —OH attached directly to carbons marked 2 and 5.

It should also be noted that tautomeric forms such as, for example, themoieties:

are considered equivalent in certain embodiments of this invention.

“Alkynylalkyl” means an alkynyl-alkyl-group in which the alkynyl andalkyl are as previously described. Preferred alkynylalkyls contain alower alkynyl and a lower alkyl group. The bond to the parent moiety isthrough the alkyl. Non-limiting examples of suitable alkynylalkyl groupsinclude propargylmethyl.

“Heteroaralkyl” means a heteroaryl-alkyl-group in which the heteroaryland alkyl are as previously described. Preferred heteroaralkyls containa lower alkyl group. Non-limiting examples of suitable aralkyl groupsinclude pyridylmethyl, and quinolin-3-ylmethyl. The bond to the parentmoiety is through the alkyl.

“Hydroxyalkyl” means a HO-alkyl-group in which alkyl is as previouslydefined. Preferred hydroxyalkyls contain lower alkyl. Non-limitingexamples of suitable hydroxyalkyl groups include hydroxymethyl and2-hydroxyethyl.

“Acyl” means an H—C(O)—, alkyl-C(O)— or cycloalkyl-C(O)—, group in whichthe various groups are as previously described. The bond to the parentmoiety is through the carbonyl. Preferred acyls contain a lower alkyl.Non-limiting examples of suitable acyl groups include formyl, acetyl andpropanoyl.

“Aroyl” means an aryl-C(O)— group in which the aryl group is aspreviously described. The bond to the parent moiety is through thecarbonyl. Non-limiting examples of suitable groups include benzoyl and1-naphthoyl.

“Alkoxy” means an alkyl-O— group in which the alkyl group is aspreviously described. Non-limiting examples of suitable alkoxy groupsinclude methoxy, ethoxy, n-propoxy, isopropoxy and n-butoxy. The bond tothe parent moiety is through the ether oxygen.

“Aryloxy” means an aryl-O— group in which the aryl group is aspreviously described. Non-limiting examples of suitable aryloxy groupsinclude phenoxy and naphthoxy. The bond to the parent moiety is throughthe ether oxygen.

“Aralkyloxy” means an aralkyl-O— group in which the aralkyl group is aspreviously described. Non-limiting examples of suitable aralkyloxygroups include benzyloxy and 1- or 2-naphthalenemethoxy. The bond to theparent moiety is through the ether oxygen.

“Alkylthio” means an alkyl-S— group in which the alkyl group is aspreviously described. Non-limiting examples of suitable alkylthio groupsinclude methylthio and ethylthio. The bond to the parent moiety isthrough the sulfur.

“Arylthio” means an aryl-S— group in which the aryl group is aspreviously described. Non-limiting examples of suitable arylthio groupsinclude phenylthio and naphthylthio. The bond to the parent moiety isthrough the sulfur.

“Aralkylthio” means an aralkyl-S— group in which the aralkyl group is aspreviously described. Non-limiting example of a suitable aralkylthiogroup is benzylthio. The bond to the parent moiety is through thesulfur.

“Alkoxycarbonyl” means an alkyl-O—CO— group. Non-limiting examples ofsuitable alkoxycarbonyl groups include methoxycarbonyl andethoxycarbonyl. The bond to the parent moiety is through the carbonyl.

“Aryloxycarbonyl” means an aryl-O—C(O)— group. Non-limiting examples ofsuitable aryloxycarbonyl groups include phenoxycarbonyl andnaphthoxycarbonyl. The bond to the parent moiety is through thecarbonyl.

“Aralkoxycarbonyl” means an aralkyl-O—C(O)— group. Non-limiting exampleof a suitable aralkoxycarbonyl group is benzyloxycarbonyl. The bond tothe parent moiety is through the carbonyl.

“Alkylsulfonyl” means an alkyl-S(O₂)— group. Preferred groups are thosein which the alkyl group is lower alkyl. The bond to the parent moietyis through the sulfonyl.

“Arylsulfonyl” means an aryl-S(O₂)— group. The bond to the parent moietyis through the sulfonyl.

The term “substituted” means that one or more hydrogens on thedesignated atom is replaced with a selection from the indicated group,provided that the designated atom's normal valency under the existingcircumstances is not exceeded, and that the substitution results in astable compound. Combinations of substituents and/or variables arepermissible only if such combinations result in stable compounds. By“stable compound’ or “stable structure” is meant a compound that issufficiently robust to survive isolation to a useful degree of purityfrom a reaction mixture, and formulation into an efficacious therapeuticagent.

The term “optionally substituted” means optional substitution with thespecified groups, radicals or moieties.

The term “isolated” or “in isolated form” for a compound refers to thephysical state of said compound after being isolated from a syntheticprocess or natural source or combination thereof. The term “purified” or“in purified form” for a compound refers to the physical state of saidcompound after being obtained from a purification process or processesdescribed herein or well known to the skilled artisan, in sufficientpurity to be characterizable by standard analytical techniques describedherein or well known to the skilled artisan.

It should also be noted that any heteroatom with unsatisfied valences inthe text, schemes, examples and Tables herein is assumed to have thehydrogen atom(s) to satisfy the valences.

When a functional group in a compound is termed “protected”, this meansthat the group is in modified form to preclude undesired side reactionsat the protected site when the compound is subjected to a reaction.Suitable protecting groups will be recognized by those with ordinaryskill in the art as well as by reference to standard textbooks such as,for example, T. W. Greene et al, Protective Groups in organic Synthesis(1991), Wiley, New York

When any variable (e.g., aryl, heterocycle, R², etc.) occurs more thanone time in any constituent or in Formula III, its definition on eachoccurrence is independent of its definition at every other occurrence.

As used herein, the term “composition” is intended to encompass aproduct comprising the specified ingredients in the specified amounts,as well as any product which results, directly or indirectly, fromcombination of the specified ingredients in the specified amounts.

Prodrugs and solvates of the compounds of the invention are alsocontemplated herein. The term “prodrug”, as employed herein, denotes acompound that is a drug precursor which, upon administration to asubject, undergoes chemical conversion by metabolic or chemicalprocesses to yield a compound of Formula III or a salt and/or solvatethereof. A discussion of prodrugs is provided in T. Higuchi and V.Stella, Pro-drugs as Novel Delivery Systems (1987) 14 of the A.C.S.Symposium Series, and in Bioreversible Carriers in Drug Design, (1987)Edward B. Roche, ed., American Pharmaceutical Association and PergamonPress, both of which are incorporated herein by reference thereto.

“Solvate” means a physical association of a compound of this inventionwith one or more solvent molecules. This physical association involvesvarying degrees of ionic and covalent bonding, including hydrogenbonding. In certain instances the solvate will be capable of isolation,for example when one or more solvent molecules are incorporated in thecrystal lattice of the crystalline solid. “Solvate” encompasses bothsolution-phase and isolatable solvates. Non-limiting examples ofsuitable solvates include ethanolates, methanolates, and the like.“Hydrate” is a solvate wherein the solvent molecule is H₂O.

“Effective amount” or “therapeutically effective amount” is meant todescribe an amount of compound or a composition of the present inventioneffective in inhibiting the CDK(s) and thus producing the desiredtherapeutic, ameliorative, inhibitory or preventative effect.

The compounds of Formula III can form salts which are also within thescope of this invention. Reference to a compound of Formula III hereinis understood to include reference to salts thereof, unless otherwiseindicated. The term “salt(s)”, as employed herein, denotes acidic saltsformed with inorganic and/or organic acids, as well as basic saltsformed with inorganic and/or organic bases. In addition, when a compoundof Formula III contains both a basic moiety, such as, but not limited toa pyridine or imidazole, and an acidic moiety, such as, but not limitedto a carboxylic acid, zwitterions (“inner salts”) may be formed and areincluded within the term “salt(s)” as used herein. Pharmaceuticallyacceptable (i.e., non-toxic, physiologically acceptable) salts arepreferred, although other salts are also useful. Salts of the compoundsof the Formula III may be formed, for example, by reacting a compound ofFormula III with an amount of acid or base, such as an equivalentamount, in a medium such as one in which the salt precipitates or in anaqueous medium followed by lyophilization.

Exemplary acid addition salts include acetates, ascorbates, benzoates,benzenesulfonates, bisulfates, borates, butyrates, citrates,camphorates, camphorsulfonates, fumarates, hydrochlorides,hydrobromides, hydroiodides, lactates, maleates, methanesulfonates,naphthalenesulfonates, nitrates, oxalates, phosphates, propionates,salicylates, succinates, sulfates, tartarates, thiocyanates,toluenesulfonates (also known as tosylates,) and the like. Additionally,acids which are generally considered suitable for the formation ofpharmaceutically useful salts from basic pharmaceutical compounds arediscussed, for example, by S. Berge et al, Journal of PharmaceuticalSciences (1977) 66(1) 1–19; P. Gould, International J. of Pharmaceutics(1986) 33 201–217; Anderson et al, The Practice of Medicinal Chemistry(1996), Academic Press, New York; and in The Orange Book (Food & DrugAdministration, Washington, D.C. on their website). These disclosuresare incorporated herein by reference thereto.

Exemplary basic salts include ammonium salts, alkali metal salts such assodium, lithium, and potassium salts, alkaline earth metal salts such ascalcium and magnesium salts, salts with organic bases (for example,organic amines) such as dicyclohexylamines, t-butyl amines, and saltswith amino acids such as arginine, lysine and the like. Basicnitrogen-containing groups may be quarternized with agents such as loweralkyl halides (e.g. methyl, ethyl, and butyl chlorides, bromides andiodides), dialkyl sulfates (e.g. dimethyl, diethyl, and dibutylsulfates), long chain halides (e.g. decyl, lauryl, and stearylchlorides, bromides and iodides), aralkyl halides (e.g. benzyl andphenethyl bromides), and others.

All such acid salts and base salts are intended to be pharmaceuticallyacceptable salts within the scope of the invention and all acid and basesalts are considered equivalent to the free forms of the correspondingcompounds for purposes of the invention.

Pharmaceutically acceptable esters of the present compounds include thefollowing groups: (1) carboxylic acid esters obtained by esterificationof the hydroxy groups, in which the non-carbonyl moiety of thecarboxylic acid portion of the ester grouping is selected from straightor branched chain alkyl (for example, acetyl, n-propyl, t-butyl, orn-butyl), alkoxyalkyl (for example, methoxymethyl), aralkyl (forexample, benzyl), aryloxyalkyl (for example, phenoxymethyl), aryl (forexample, phenyl optionally substituted with, for example, halogen,C₁₋₄alkyl, or C₁₋₄alkoxy or amino); (2) sulfonate esters, such as alkyl-or aralkylsulfonyl (for example, methanesulfonyl); (3) amino acid esters(for example, L-valyl or L-isoleucyl); (4) phosphonate esters and (5)mono-, di- or triphosphate esters. The phosphate esters may be furtheresterified by, for example, a C₁₋₂₀ alcohol or reactive derivativethereof, or by a 2,3-di (C₆₋₂₄)acyl glycerol.

Compounds of Formula III, and salts, solvates and prodrugs thereof, mayexist in their tautomeric form (for example, as an amide or iminoether). All such tautomeric forms are contemplated herein as part of thepresent invention.

All stereoisomers (for example, geometric isomers, optical isomers andthe like) of the present compounds (including those of the salts,solvates and prodrugs of the compounds as well as the salts and solvatesof the prodrugs), such as those which may exist due to asymmetriccarbons on various substituents, including enantiomeric forms (which mayexist even in the absence of asymmetric carbons), rotameric forms,atropisomers, and diastereomeric forms, are contemplated within thescope of this invention, as are positional isomers (such as, forexample, 4-pyridyl and 3-pyridyl). Individual stereoisomers of thecompounds of the invention may, for example, be substantially free ofother isomers, or may be admixed, for example, as racemates or with allother, or other selected, stereoisomers. The chiral centers of thepresent invention can have the S or R configuration as defined by theIUPAC 1974 Recommendations. The use of the terms “salt”, “solvate”“prodrug” and the like, is intended to equally apply to the salt,solvate and prodrug of enantiomers, stereoisomers, rotamers, tautomers,positional isomers, racemates or prodrugs of the inventive compounds.

Polymorphic forms of the compounds of Formula III, and of the salts,solvates and prodrugs of the compounds of Formula III, are intended tobe included in the present invention.

The compounds according to the invention have pharmacologicalproperties; in particular, the compounds of Formula III can beinhibitors of protein kinases such as, for example, the inhibitors ofthe cyclin-dependent kinases (e.g., CDK1, CDK2, CDK3, CDK4 CDK5 and thelike), mitogen-activated protein kinase (MAPK/ERK), glycogen synthasekinase 3(GSK3beta), checkpoint kinase-1 (“CHK-1”), checkpoint kinase-2(“CHK-2”), Aurora kinases, (e.g., Aurora A, B, and C), protein kinase B(e.g., serine/threonine kinases such as AKT1, AKT2 and AKT3), and thelike. The cyclin dependent kinases (CDKs) include, for example, CDC2(CDK1), CDK2, CDK4, CDK5, CDK6, CDK7 and CDK8. The novel compounds ofFormula III are expected to be useful in the therapy of proliferativediseases such as cancer, autoimmune diseases, viral diseases, fungaldiseases, neurological/neurodegenerative disorders, arthritis,inflammation, anti-proliferative (e.g., ocular retinopathy), neuronal,alopecia and cardiovascular disease. Many of these diseases anddisorders are listed in U.S. Pat. No. 6,413,974 cited earlier, thedisclosure of which is incorporated herein.

More specifically, the compounds of Formula III can be useful in thetreatment of a variety of cancers, including (but not limited to) thefollowing: carcinoma, including that of the bladder, breast, colon,kidney, liver, lung, including small cell lung cancer, esophagus, gallbladder, ovary, pancreas, stomach, cervix, thyroid, prostate, and skin,including squamous cell carcinoma;

hematopoietic tumors of lymphoid lineage, including leukemia, acutelymphocytic leukemia, acute lymphoblastic leukemia, B-cell lymphoma,T-cell lymphoma, Hodgkins lymphoma, non-Hodgkins lymphoma, hairy celllymphoma and Burkett's lymphoma;

hematopoietic tumors of myeloid lineage, including acute and chronicmyelogenous leukemias, myelodysplastic syndrome and promyelocyticleukemia;

tumors of mesenchymal origin, including fibrosarcoma andrhabdomyosarcoma;

tumors of the central and peripheral nervous system, includingastrocytoma, neuroblastoma, glioma and schwannomas; and

other tumors, including melanoma, seminoma, teratocarcinoma,osteosarcoma, xenoderoma pigmentosum, keratoctanthoma, thyroidfollicular cancer and Kaposi's sarcoma.

Due to the key role of CDKs in the regulation of cellular proliferationin general, inhibitors could act as reversible cytostatic agents whichmay be useful in the treatment of any disease process which featuresabnormal cellular proliferation, e.g., benign prostate hyperplasia,familial adenomatosis polyposis, neuro-fibromatosis, atherosclerosis,pulmonary fibrosis, arthritis, psoriasis, glomerulonephritis, restenosisfollowing angioplasty or vascular surgery, hypertrophic scar formation,inflammatory bowel disease, transplantation rejection, endotoxic shock,and fungal infections.

Compounds of Formula III may also be useful in the treatment ofAlzheimer's disease, as suggested by the recent finding that CDK5 isinvolved in the phosphorylation of tau protein (J. Biochem, (1995) 117,741–749).

Compounds of Formula III may induce or inhibit apoptosis. The apoptoticresponse is aberrant in a variety of human diseases. Compounds ofFormula III, as modulators of apoptosis, will be useful in the treatmentof cancer (including but not limited to those types mentionedhereinabove), viral infections (including but not limited to herpevirus,poxvirus, Epstein-Barr virus, Sindbis virus and adenovirus), preventionof AIDS development in HIV-infected individuals, autoimmune diseases(including but not limited to systemic lupus, erythematosus, autoimmunemediated glomerulonephritis, rheumatoid arthritis, psoriasis,inflammatory bowel disease, and autoimmune diabetes mellitus),neurodegenerative disorders (including but not limited to Alzheimer'sdisease, AIDS-related dementia, Parkinson's disease, amyotrophic lateralsclerosis, retinitis pigmentosa, spinal muscular atrophy and cerebellardegeneration), myelodysplastic syndromes, aplastic anemia, ischemicinjury associated with myocardial infarctions, stroke and reperfusioninjury, arrhythmia, atherosclerosis, toxin-induced or alcohol relatedliver diseases, hematological diseases (including but not limited tochronic anemia and aplastic anemia), degenerative diseases of themusculoskeletal system (including but not limited to osteoporosis andarthritis) aspirin-sensitive rhinosinusitis, cystic fibrosis, multiplesclerosis, kidney diseases and cancer pain.

Compounds of Formula III, as inhibitors of the CDKs, can modulate thelevel of cellular RNA and DNA synthesis. These agents would therefore beuseful in the treatment of viral infections (including but not limitedto HIV, human papilloma virus, herpesvirus, poxvirus, Epstein-Barrvirus, Sindbis virus and adenovirus).

Compounds of Formula III may also be useful in the chemoprevention ofcancer. Chemoprevention is defined as inhibiting the development ofinvasive cancer by either blocking the initiating mutagenic event or byblocking the progression of pre-malignant cells that have alreadysuffered an insult or inhibiting tumor relapse.

Compounds of Formula III may also be useful in inhibiting tumorangiogenesis and metastasis.

Compounds of Formula III may also act as inhibitors of other proteinkinases, e.g., protein kinase C, her2, raf 1, MEK1, MAP kinase, EGFreceptor, PDGF receptor, IGF receptor, PI3 kinase, wee1 kinase, Src, Abland thus be effective in the treatment of diseases associated with otherprotein kinases.

Another aspect of this invention is a method of treating a mammal (e.g.,human) having a disease or condition associated with the CDKs byadministering a therapeutically effective amount of at least onecompound of Formula III, or a pharmaceutically acceptable salt, solvateor ester of said compound to the mammal.

A preferred dosage is about 0.001 to 500 mg/kg of body weight/day of thecompound of Formula III. An especially preferred dosage is about 0.01 to25 mg/kg of body weight/day of a compound of Formula III, or apharmaceutically acceptable salt, solvate or ester of said compound.

The compounds of this invention may also be useful in combination(administered together or sequentially) with one or more of anti-cancertreatments such as radiation therapy, and/or one or more anti-canceragents selected from the group consisting of cytostatic agents,cytotoxic agents (such as for example, but not limited to, DNAinteractive agents (such as cisplatin or doxorubicin)); taxanes (e.g.taxotere, taxol); topoisomerase II inhibitors (such as etoposide);topoisomerase I inhibitors (such as irinotecan (or CPT-11), camptostar,or topotecan); tubulin interacting agents (such as paclitaxel, docetaxelor the epothilones); hormonal agents (such as tamoxifen); thymidilatesynthase inhibitors (such as 5-fluorouracil); anti-metabolites (such asmethoxtrexate); alkylating agents (such as temozolomide (TEMODAR™ fromSchering-Plough Corporation, Kenilworth, N.J.), cyclophosphamide);Farnesyl protein transferase inhibitors (such as,SARASAR™(4-[2-[4-[(11R)-3,10-dibromo-8-chloro-6,11-dihydro-5H-benzo[5,6]cyclohepta[1,2-b]pyridin-11-yl-]-1-piperidinyl]-2-oxoehtyl]-1-piperidinecarboxamide, or SCH 66336from Schering-Plough Corporation, Kenilworth, N.J.), tipifarnib(Zarnestra® or R115777 from Janssen Pharmaceuticals), L778,123 (afarnesyl protein transferase inhibitor from Merck & Company, WhitehouseStation, N.J.), BMS 214662 (a farnesyl protein transferase inhibitorfrom Bristol-Myers Squibb Pharmaceuticals, Princeton, N.J.); signaltransduction inhibitors (such as, Iressa (from Astra ZenecaPharmaceuticals, England), Tarceva (EGFR kinase inhibitors), antibodiesto EGFR (e.g., C225), GLEEVEC™ (C-abl kinase inhibitor from NovartisPharmaceuticals, East Hanover, N.J.); interferons such as, for example,intron (from Schering-Plough Corporation), Peg-Intron (fromSchering-Plough Corporation); hormonal therapy combinations; aromatasecombinations; ara-C, adriamycin, cytoxan, and gemcitabine.

Other anti-cancer (also known as anti-neoplastic) agents include but arenot limited to Uracil mustard, Chlormethine, Ifosfamide, Melphalan,Chlorambucil, Pipobroman, Triethylenemelamine,Triethylenethiophosphoramine, Busulfan, Carmustine, Lomustine,Streptozocin, Dacarbazine, Floxuridine, Cytarabine, 6-Mercaptopurine,6-Thioguanine, Fludarabine phosphate, oxaliplatin, leucovirin,oxaliplatin (ELOXATIN™ from Sanofi-Synthelabo Pharmaeuticals, France),Pentostatine, Vinblastine, Vincristine, Vindesine, Bleomycin,Dactinomycin, Daunorubicin, Doxorubicin, Epirubicin, Idarubicin,Mithramycin, Deoxycoformycin, Mitomycin-C, L-Asparaginase, Teniposide17α-Ethinylestradiol, Diethylstilbestrol, Testosterone, Prednisone,Fluoxymesterone, Dromostanolone propionate, Testolactone,Megestrolacetate, Methylprednisolone, Methyltestosterone, Prednisolone,Triamcinolone, Chlorotrianisene, Hydroxyprogesterone, Aminoglutethimide,Estramustine, Medroxyprogesteroneacetate, Leuprolide, Flutamide,Toremifene, goserelin, Cisplatin, Carboplatin, Hydroxyurea, Amsacrine,Procarbazine, Mitotane, Mitoxantrone, Levamisole, Navelbene,Anastrazole, Letrazole, Capecitabine, Reloxafine, Droloxafine, orHexamethylmelamine.

When administering a combination therapy to a patient in need of suchadministration, the therapeutic agents in the combination, or apharmaceutical composition or compositions comprising the therapeuticagents, may be administered in any order such as, for example,sequentially, concurrently, together, simultaneously and the like. Theamounts of the various actives in such combination therapy may bedifferent amounts (different dosage amounts) or same amounts (samedosage amounts). Thus, for non-limiting illustration purposes, acompound of Formula III and an additional therapeutic agent may bepresent in fixed amounts (dosage amounts) in a single dosage unit (e.g.,a capsule, a tablet and the like). A commercial example of such singledosage unit containing fixed amounts of two different active compoundsis VYTORIN® (available from Merck Schering-Plough Pharmaceuticals,Kenilworth, N.J.).

If formulated as a fixed dose, such combination products employ thecompounds of this invention within the dosage range described herein andthe other pharmaceutically active agent or treatment within its dosagerange. For example, the CDC2 inhibitor olomucine has been found to actsynergistically with known cytotoxic agents in inducing apoptosis (J.Cell Sci., (1995) 108, 2897. Compounds of Formula III may also beadministered sequentially with known anticancer or cytotoxic agents whena combination formulation is inappropriate. The invention is not limitedin the sequence of administration; compounds of Formula III may beadministered either prior to or after administration of the knownanticancer or cytotoxic agent. For example, the cytotoxic activity ofthe cyclin-dependent kinase inhibitor flavopiridol is affected by thesequence of administration with anticancer agents, Cancer Research,(1997) 57, 3375. Such techniques are within the skills of personsskilled in the art as well as attending physicians.

Accordingly, in an aspect, this invention includes combinationscomprising an amount of at least one compound of Formula III, or apharmaceutically acceptable salt, solvate or ester thereof, and anamount of one or more anti-cancer treatments and anti-cancer agentslisted above wherein the amounts of the compounds/treatments result indesired therapeutic effect.

The pharmacological properties of the compounds of this invention may beconfirmed by a number of pharmacological assays. The exemplifiedpharmacological assays which are described later can be carried out withthe compounds according to the invention and their salts.

This invention is also directed to pharmaceutical compositions whichcomprise at least one compound of Formula III, or a pharmaceuticallyacceptable salt, solvate or ester of said compound and at least onepharmaceutically acceptable carrier.

For preparing pharmaceutical compositions from the compounds describedby this invention, inert, pharmaceutically acceptable carriers can beeither solid or liquid. Solid form preparations include powders,tablets, dispersible granules, capsules, cachets and suppositories. Thepowders and tablets may be comprised of from about 5 to about 95 percentactive ingredient. Suitable solid carriers are known in the art, e.g.,magnesium carbonate, magnesium stearate, talc, sugar or lactose.Tablets, powders, cachets and capsules can be used as solid dosage formssuitable for oral administration. Examples of pharmaceuticallyacceptable carriers and methods of manufacture for various compositionsmay be found in A. Gennaro (ed.), Remington's Pharmaceutical Sciences,18^(th) Edition, (1990), Mack Publishing Co., Easton, Pa.

Liquid form preparations include solutions, suspensions and emulsions.As an example may be mentioned water or water-propylene glycol solutionsfor parenteral injection or addition of sweeteners and opacifiers fororal solutions, suspensions and emulsions. Liquid form preparations mayalso include solutions for intranasal administration.

Aerosol preparations suitable for inhalation may include solutions andsolids in powder form, which may be in combination with apharmaceutically acceptable carrier, such as an inert compressed gas,e.g. nitrogen.

Also included are solid form preparations that are intended to beconverted, shortly before use, to liquid form preparations for eitheroral or parenteral administration. Such liquid forms include solutions,suspensions and emulsions.

The compounds of the invention may also be deliverable transdermally.The transdermal compositions can take the form of creams, lotions,aerosols and/or emulsions and can be included in a transdermal patch ofthe matrix or reservoir type as are conventional in the art for thispurpose.

The compounds of this invention may also be delivered subcutaneously.

Preferably the compound is administered orally or intravenously.

Preferably, the pharmaceutical preparation is in a unit dosage form. Insuch form, the preparation is subdivided into suitably sized unit dosescontaining appropriate quantities of the active component, e.g., aneffective amount to achieve the desired purpose.

The quantity of active compound in a unit dose of preparation may bevaried or adjusted from about 1 mg to about 100 mg, preferably fromabout 1 mg to about 50 mg, more preferably from about 1 mg to about 25mg, according to the particular application.

The actual dosage employed may be varied depending upon the requirementsof the patient and the severity of the condition being treated.Determination of the proper dosage regimen for a particular situation iswithin the skill of the art. For convenience, the total daily dosage maybe divided and administered in portions during the day as required.

The amount and frequency of administration of the compounds of theinvention and/or the pharmaceutically acceptable salts thereof will beregulated according to the judgment of the attending clinicianconsidering such factors as age, condition and size of the patient aswell as severity of the symptoms being treated. A typical recommendeddaily dosage regimen for oral administration can range from about 1mg/day to about 500 mg/day, preferably 1 mg/day to 200 mg/day, in two tofour divided doses.

Another aspect of this invention is a kit comprising a therapeuticallyeffective amount of at least one compound of Formula III, or apharmaceutically acceptable salt, solvate or ester of said compound anda pharmaceutically acceptable carrier, vehicle or diluent.

Yet another aspect of this invention is a kit comprising an amount of atleast one compound of Formula III, or a pharmaceutically acceptablesalt, solvate or ester of said compound and an amount of at least oneanticancer therapy and/or anti-cancer agent listed above, wherein theamounts of the two or more ingredients result in desired therapeuticeffect.

The invention disclosed herein is exemplified by the followingpreparations, proposed pathways and examples which should not beconstrued to limit the scope of the disclosure. Alternative mechanisticpathways and analogous structures will be apparent to those skilled inthe art.

Where NMR data are presented, ¹H spectra were obtained on either aVarian VXR-200 (200 MHz, ¹H), Varian Gemini-300 (300 MHz) or XL400 (400MHz) and are reported as ppm down field from Me₄Si with number ofprotons, multiplicities, and coupling constants in Hertz indicatedparenthetically. Where LC/MS data are presented, analyses was performedusing an Applied Biosystems API-100 mass spectrometer and ShimadzuSCL-10A LC column: Altech platinum C18, 3 micron, 33 mm×7 mm ID;gradient flow: 0 min—10% CH₃CN, 5 min—95% CH₃CN, 7 min—95% CH₃CN, 7.5min—10% CH₃CN, 9 min—stop, and the retention time and observed parention are given.

The following solvents and reagents may be referred to by theirabbreviations:

Thin layer chromatography: TLC dichloromethane: CH₂Cl₂ ethyl acetate:AcOEt or EtOAc methanol: MeOH trifluoroacetate: TFA triethylamine: Et₃Nor TEA butoxycarbonyl: n-Boc or Boc nuclear magnetic resonancespectroscopy: NMR liquid chromatography mass spectrometry: LCMS highresolution mass spectrometry: HRMS milliliters: mL millimoles: mmolmicroliters: μl grams: g milligrams: mg room temperature or rt(ambient): about 25° C. dimethoxyethane: DME

EXAMPLES

In general the compounds described in this invention can be prepared asillustrated in Scheme 1. Acylation of the appropriately substituted3-aminopyrazole 1 and base-catalyzed cyclization leads to thiol 2.

Methylation, chlorination, and displacement with the desired amine gives4, which can be transformed into the desired products 5 by oxidation andnucleophilic displacement.

Preparative Example 100

To a solution of 4-aminomethylpyridine (1.41 mL, 13.87 mmol) in CH₂Cl₂(50 mL) was added BOC₂O (3.3 g, 1.1 eq.) and TEA and the resultingsolution was stirred a room temperature 2 hours. The reaction mixturewas diluted with H₂O (50 mL) and extracted with CH₂Cl₂. The combinedorganics were dried over Na₂SO₄, filtered and concentrated under reducedpressure. The crude product was purified by flash chromatography using a5% (10% NH₄OH in MeOH) solution in CH₂Cl₂ as eluent to give a yellowsolid (2.62 g, 91% yield). LCMS: MH⁺=209.

Preparative Example 101

By essentially the same procedure set forth in Preparative Example 100only substituting 3-aminomethylpyridine, the above compound was preparedas a yellow oil (2.66 g, 92% yield). LCMS: MH⁺=209.

Preparative Example 200

To a solution of the compound prepared in Preparative Example 100 (0.20g, 0.96 mmol) in CH₂Cl₂ (5 mL) at 0° C. was added m-CPBA (0.17 g, 1.0eq) and the resulting solution stirred at 0° C. 2 hours and stored at 4°C. overnight at which time the reaction mixture was warmed to roomtemperature and stirred 3 hours. The reaction mixture was diluted withH₂O and extracted with CH₂Cl₂. The combined organics were dried overNa₂SO₄, filtered, and concentrated. The crude product was purified byflash chromatography using a 10% (10% NH₄OH in MeOH) solution as eluent:LCMS: MH⁺=255.

Preparative Example 201

A solution of oxone (58.6 g) in H₂O (250 mL) was added dropwise to thecompound prepared in Preparative Example 101 (27 g, 0.13 mol) and NaHCO₃(21.8 g, 2.0 eq.) in MeOH (200 mL) and H₂O (250 mL). The resultingsolution was stirred at room temperature overnight. The reaction mixturewas diluted with CH2Cl2 (500 mL) and filtered. The layers were separatedand the aqueous layer extracted with CH₂Cl₂. The combined organics weredried over Na₂SO₄, filtered, and concentrated under reduced pressure togive a white solid (21.0 g, 72% yield). MS: MH⁺=255.

Preparative Example 300

The compound prepared in Preparative Example 200 (0.29 g, 1.29 mmol) wasstirred at room temperature in 4M HCl in dioxane (0.97 mL) 2 hours. Thereaction mixture was concentrated in vacuo and used without furtherpurification. LCMS: MH⁺=125.

Preparative Example 301

By essentially the same procedure set forth in Preparative Example 201only substituting the compound prepared in Preparative Example 201, thecompound shown above was prepared. LCMS: MH⁺=125.

Preparative Example 400

Step A:

A solution of aldehyde (50 g, 0.41 mol) [see, for example, WO 0232893]in MeOH (300 mL) was cooled to 0° C. and carefully treated with NaBH₄(20 g, 0.53 mol in 6 batches) over 20 minutes. The reaction was thenallowed to warm to 20° C. and was stirred for 4 hours. The mixture wasagain cooled to 0° C., carefully quenched with saturated aqueous NH₄Cl,and concentrated. Flash chromatography (5–10% 7N NH₃—MeOH/CH₂Cl₂)provided the primary alcohol (31 g, 62%) as a light yellow solid.

Step B:

A slurry of alcohol (31 g, 0.25 mol) from Preparative Example 400, StepA in CH₂Cl₂ (500 mL) was cooled to 0° C. and slowly treated with SOCl₂(55 mL, 0.74 mol over 30 minutes). The reaction was then stirredovernight at 20° C. The material was concentrated, slurried in acetone,and then filtered. The resulting beige solid was dried overnight invacuo (38.4 g, 52%, HCl salt).

Step C:

To a 15 mL pressure tube charged with a stir bar was added chloride (150mg, 0.83 mmol) from Preparative Example 400, Step B followed by 7 MNH₃/MeOH (10 mL). The resulting solution was stirred for 48 h at rtwhereupon the mixture was concentrated under reduced pressure to afforda light yellow solid (0.146 g, 83%). M+H (free base)=140.

Preparative Example 401

The known primary alcohol was prepared according to WO 00/37473 and wasconverted to the desired amine dihydrochloride in analogous fashion asPreparative Example 400 according to WO 02/064211.

Preparative Example 500

Piperidine-2-ethanol (127 g, 980 mmol) in 95% EtOH (260 mL) was added to(S)-(+)-camphorsulfonic acid (228.7 g, 1.0 eq.) in 95% EtOH (150 mL) andthe resulting solution was warmed to reflux. To the warm solution wasadded Et₂O (600 mL) and the solution cooled to room temperature and letstand 3 days. The resulting crystals were filtered and dried in vacuo(25 g): mp 173–173° C. (lit. 168° C.). The salt was then dissolved inNaOH (3M, 100 mL) and stirred 2 hours and the resulting solution wasextracted with CH₂Cl₂ (5×100 mL). The combined organics were dried overNa₂SO₄, filtered, filtered and concentrated under reduced pressure togive (S)-piperidine-2-ethanol (7.8 g) a portion of which wasrecrystallized from Et₂O: mp=69–70° C. (lit. 68–69° C.); [α]_(D)=14.090(CHCl₃, c=0.2).

Preparative Example 501

Bye essentially the same procedure set forth in Preparative Example 500only substituting (R)-(−)-camphorsulfonic acid, (R)-piperidine-2-ethanolwas prepared. (1.27 g): [α]_(D)=11.3° (CHCl₃, c=0.2).

Preparative Example 502

To pressure bottle charged with a solution ofcis-(1R,2S)-(+)-2-(Benzylamino)cyclohexanemethanol (1g, 4.57 mmol) inMeOH (35 mL) was added 20% wt Pd(OH)₂ (0.3 g, >50% wet) in one portion.The mixture was shaken under 50 psi of H₂ in a Parr hydrogenationapparatus for 12 h. The mixture was purged to N₂ and was filteredthrough a pad of Celite. The pad was generously washed with MeOH (2×25mL) and the resulting filtrate was concentrated under reduces pressureto afford 0.57 g (97%) of a white solid. M+H=130.

Preparative Example 507

t-BuOK (112.0 g, 1.00 mol) was stirred under N₂ in dry Et₂O (3.0 L) in a5 L flask equipped with an addition funnel. A mixture of butyronitrile(69.0 g, 1.00 mol) and ethylformate (77.7 g, 1.05 mol) was addeddropwise during 3 hrs, the reaction mixture was then stirred overnightat room temperature. The mixture was cooled to 0° C., AcOH (57 mL) wasadded, the mixture was filtered, and the solid was washed with Et₂O (500mL). The combined filtrates were evaporated at room temperature on arotovap to give pale yellow oil (95.1 g).

The oil was dissolved in dry EtOH (100 mL), 99% hydrazine monohydrate(48 mL) was added, then AcOH (14 mL) was added, and the mixture wasrefluxed under N₂ overnight. The solvents were evaporated and theresulting oil was chromatographed on silica gel with CH₂Cl₂:7N NH₃ inMeOH. 22.4 g (20%) of 3-amino-4-ethylpyrazole was obtained as clear oilthat solidified upon standing.

Preparative Example 508

Prep. Ex. Column 2 508

By essentially the same procedure set forth in Preparative Example 507only substituting the appropriate starting materials, the aminopyrazole508 was prepared.

Preparative Example 509–511

By essentially the same procedure set forth in Preparative Example 507only substituting the appropriate starting materials, the aminopyrazolesshown in Column 2 of Table 500 are prepared.

TABLE 500 Prep. Ex. Column 2 509

510

511

Preparative Example 512

Step A:

To a strirred solution of the pyrazole (3.33 g, 30 0 mmol) fromPreparative Example 507 in dry CH₂Cl₂ (50 mL) at 0° C. was dropwiseadded ethoxycarbonyl isothiocyanate (3.54 mL, 3.00 mmol). The resultingmixture was stirred at room temperature 24 hours at which time theprecipitate was filtered, washed with Et₂O (2×50 mL), and dried in avacuum. White solid (3.50 g, 48%) was obtained. LCMS: MH⁺=243.Mp=177–179° C.

Step B:

A mixture of the solid from Preparative Example 512, Step A (800 mg,3.30 mmol) and K₂CO₃ (1.37 g, 9.90 mmol) in dry acetonitrile (20 mL) wasstirred and refluxed under nitrogen for 4 hours. The mixture was cooledto 25° C., acidified with acetic acid (5 mL), and diluted with water (20mL). The solvents were evaporated and the residue was suspended in water(50 mL). The solid was filtered off, washed on filter with water (2×20mL), and dried in a vacuum. An off white solid (548 mg, 85%) wasobtained. LCMS: MH⁺=197. Mp=251–253° C.

Preparative Example 513

Step A:

To a solution of the compound described in Preparative Example 512 (500mg, 2.55 mmol) in EtOH was added NaOH (204 mg, 5.10 mmol) in H₂O (3 mL)and then Mel (362 mg, 2.55 mmol) dropwise. The resulting mixture wasstirred at room temperature 1 hour, acidified with 1 M HCl (5 mL), andthe solvents were evaporated. The residue was purified by columnchromatography on silicagel with CH₂Cl₂/MeOH (15:1) as eluent to yield awhite solid (442 mg, 83%). LCMS: MH⁺=211. Mp=182–184° C.

Step B:

To a solution of the compound described in Preparative 513, Step A (400mg, 1.90 mmol) in POCl₃ (6 mL) was added N,N-dimethyl aniline (460 mg,3.80 mmol) and the mixture was heated to reflux under nitrogen for 18hours. The resulting solution was poured over crushed ice (200 g) andextracted with CH₂Cl₂ (2×50 mL). The combined extracts were washed withH₂O (100 mL), dried over Na2SO4, filtered, and the solvent wasevaporated. The residue was purified by column chromatography onsilicagel with CH₂Cl₂ as eluent to yield a pale yellow solid (275 mg,63%). LCMS: M⁺=229.

Preparative Example 514

Prep. Ex. Column 2 Column 3 514

By essentially the same procedures set forth in Preparative Examples512–513, compound given in Preparative Example 514 was prepared.

Preparative Example 515–517

By essentially the same procedures set forth in Preparative Examples512–513, only substituting the compounds shown in Column 2 of Table 501,the compounds shown in Column 3 of Table 501 are prepared.

TABLE 501 Prep. Ex. Column 2 Column 3 515

516

517

Preparative Example 518

A mixture of the compound from Preparative Example 513 (130 mg, 0.57mmol), the amine (94 mg, 0.68 mmol), and NaHCO3 (96 mg, 1.14 mmol) indry acetonitrile (3 mL) was stirred at 70° C. for 20 hours under N₂. Thesolvents were removed under reduced pressure and the residue waspurified by column chromatography on silicagel with CH₂Cl₂/MeOH (20:1)as eluent to yield a white solid (150 mg, 80%). LCMS: MH⁺=331.Mp=162–164° C.

Preparative Examples 519–527

By essentially the same procedure set forth in Preparative Example 518only substituting the amines in Column 3 of Table 502 and the chloridesshown in Column 3 of Table 502, the compounds shown in Column 4 of Table502 were prepared.

TABLE 502 Prep. Ex. Column 2 Column 3 Column 4 519

520

521

522

523

524

525

526

527

Example 1000

Step A:

To a solution of the compound from Preparative 518 (80 mg, 0.24 mmol) inCH₂Cl₂ (5 mL) was added 70% m-CPBA (60 mg, 0.24 mmol). The resultingsolution was stirred at room temperature overnight, then additionalCH₂Cl₂ (15 mL) was added. The solution was washed with aqueous saturatedNaHCO₃ (2×20 mL), dried over MgSO₄, filtered, and the solvent wasevaporated. So obtained white solid (55 mg) was used directly for stepB.

Step B:

A mixture of the compound from Example 1000, Step A (55 mg) with theaminoalcohol from Preparative Example 500 (60 mg) in dry NMP (0.2 mL)was stirred under N₂ at 100° C. for 5 hr. The NMP was removed underreduced pressure and the residue is purified by preparative TLC onsilicagel with CH₂Cl₂/MeOH (5:1) as eluent to yield a pale yellow waxy(30 mg, 51%). LCMS: MH⁺=412.

Examples 1001–1010

By essentially the same procedure set forth in Example 1000 onlysubstituting the appropriate amine in Column 2 of Table 1000 and theappropriate thioether in Column 3 of Table 1000 the compounds in column4 of Table 1000 were prepared.

TABLE 1000 1001

LCMS:MH⁺ = 383.1wax 1002

LCMS:MH⁺ = 398.2mp = 62–67 1003

LCMS:MH⁺ = 426.2wax 1004

LCMS:MH⁺ = 424.2gummy solid 1005

LCMS:MH⁺ = 459.1wax 1006

LCMS:MH⁺ = 445.1138–142 1007

LCMS:MH⁺ = 383.1wax 1008

LCMS:MH⁺ = 382.1wax 1009

LCMS:MH⁺ = 394.2wax 1010

LCMS:MH⁺ = 412.1wax

Examples 2001–2029

By essentially the same procedure set forth in Example 1000 onlysubstituting the appropriate amine in Column 2 of Table 2000 and theappropriate thioether in Column 3 of Table 2000 the compounds in column4 of Table 2000 can be prepared.

TABLE 2000 Ex. Column 2 Column 3 Column 4 2001

2002

2003

2004

2005

2006

2007

2008

2009

2010

2011

2012

2013

2014

2015

2016

2017

2018

2019

2020

2021

2022

2023

2024

2025

2026

2027

2028

2029

Assay

A useful assay for kinase activity is described below.

BACULOVIRUS CONSTRUCTIONS: Cyclins A and E are cloned into pFASTBAC(Invitrogen) by PCR, with the addition of a GluTAG sequence (EYMPME) atthe amino-terminal end to allow purification on anti-GluTAG affinitycolumns. The expressed proteins are approximately 46 kDa (cyclin E) and50 kDa (cyclin A) in size. CDK2 is also cloned into pFASTBAC by PCR,with the addition of a haemaglutinin epitope tag at the carboxy-terminalend (YDVPDYAS). The expressed protein is approximately 34 kDa in size.

ENZYME PRODUCTION: Recombinant baculoviruses expressing cyclins A, E andCDK2 are infected into SF9 cells at a multiplicity of infection (MOl) of5, for 48 hrs. Cells are harvested by centrifugation at 1000 RPM for 10minutes. Cyclin-containing (E or A) pellets are combined with CDK2containing cell pellets and lysed on ice for 30 minutes in five timesthe pellet volume of lysis buffer containing 50 mM Tris pH 8.0, 0.5%NP40, 1 mM DTT and protease/phosphatase inhibitors (Roche DiagnosticsGmbH, Mannheim, Germany). Mixtures are stirred for 30–60 minutes topromote cyclin-CDK2 complex formation. Mixed lysates are then spun downat 15000 RPM for 10 minutes and the supernatant retained. 5ml ofanti-GluTAG beads (for one liter of SF9 cells) are then used to capturecyclin-CDK2 complexes. Bound beads are washed three times in lysisbuffer. Proteins are competitively eluted with lysis buffer containing100–200 ug/mL of the GluTAG peptide. Eluate is dialyzed overnight in 2liters of kinase buffer containing 50 mM Tris pH 8.0, 1 mM DTT, 10 mMMgCl2, 100 uM sodium orthovanadate and 20% glycerol. Enzyme is stored inaliquots at −70° C.

IN VITRO KINASE ASSAY: CDK2 kinase assays (either cyclin A orE-dependent) are performed in low protein binding 96-well plates(Corning Inc, Corning, N.Y.). Enzyme is diluted to a final concentrationof 50 μg/ml in kinase buffer containing 50 mM Tris pH 8.0, 10 mM MgCl₂,1 mM DTT, and 0.1 mM sodium orthovanadate. The substrate used in thesereactions is a biotinylated peptide derived from Histone H1 (fromAmersham, UK). The substrate is thawed on ice and diluted to 2 μM inkinase buffer. Compounds were diluted in 10% DMSO to desirableconcentrations. For each kinase reaction, 20 μl of the 50 μg/ml enzymesolution (1 μg of enzyme) and 20 μl of the 1 μM substrate solution aremixed, then combined with 10 μl of diluted compound in each well fortesting. The kinase reaction is started by addition of 50 μl of 4 μM ATPand 1 μCi of 33P-ATP (from Amersham, UK). The reaction is allowed to runfor 1 hour at room temperature. The reaction is stopped by adding 200 μlof stop buffer containing 0.1% Triton X-100, 1 mM ATP, 5 mM EDTA, and 5mg/ml streptavidine coated SPA beads (from Amersham, UK) for 15 minutes.The SPA beads are then captured onto a 96-well GF/B filter plate(Packard/Perkin Elmer Life Sciences) using a Filtermate universalharvester (Packard/Perkin Elmer Life Sciences.). Non-specific signalsare eliminated by washing the beads twice with 2M NaCl then twice with 2M NaCl with 1% phosphoric acid. The radioactive signal is then measuredusing a TopCount 96 well liquid scintillation counter (fromPackard/Perkin Elmer Life Sciences).

IC₅₀ DETERMINATION: Dose-response curves are plotted from inhibitiondata generated, each in duplicate, from 8 point serial dilutions ofinhibitory compounds. Concentration of compound is plotted against %kinase activity, calculated by CPM of treated samples divided by CPM ofuntreated samples. To generate IC₅₀ values, the dose-response curves arethen fitted to a standard sigmoidal curve and IC₅₀ values are derived bynonlinear regression analysis. Kinase activities can be generated byusing cyclin A or cyclin E using the above-described assay. The IC₅₀ ofsome of the inventive compounds is shown below in Table 2:

TABLE 2

CDK 2 IC₅₀ [μM]0.00056

0.0013 

0.00052

0.0031 

0.106 

0.0012 

0.00088

0.00048

While the present invention has been described with in conjunction withthe specific embodiments set forth above, many alternatives,modifications and other variations thereof will be apparent to those ofordinary skill in the art. All such alternatives, modifications andvariations are intended to fall within the spirit and scope of thepresent invention.

1. A pharmaceutical composition comprising a therapeutically effectiveamount of at least one compound of the following formula, or apharmaceutically acceptable salt, or ester of said compound, incombination with at least one pharmaceutically acceptable carrier;

wherein: R¹ is selected from the group consisting of H, alkyl, aryl,heteroaryl, heteroarylalkyl, arylalkyl, NR⁶R⁷, cycloalkyl andcycloalkylalkyl, wherein each of said alkyl, aryl, heteroaryl,heteroarylalkyl, cycloalkyl, cycloalkylalkyl and arylalkyl can beunsubstituted or optionally independently substituted with one or moremoieties which can be the same or different, each being independentlyselected from the group consisting of halo, alkyl, aryl, heteroaryl,heterocyclyl, trifluoromethyl, OR⁶, NR⁶R⁷, SR⁶, SO₂R⁶, CN, SO₂N(R⁶R⁷)and NO₂; R² is alkyl, cycloalkyl, alkenyl, alkynyl, trifluoromethyl,—OR⁷, —SR⁷, hydroxyalkyl, haloalkyl, aryl, heteroaryl, halo, CN, formyl,nitro, alkylcarbonyl, aralkylcarbonyl, heteroaralkylcarbonyl, or-alkylene-N(R⁶R⁹) (where R⁸ and R⁹ independently represent H or alkyl,or R⁸ and R⁹ taken together with the nitrogen in —N(R⁸R⁹) form a five-to seven- membered heterocycle);

alkyl, alkylthio, aralkylthio, alkylsulfinyl, or aralkylsulfinyl; R⁴ isalkyl, cycloalkyl or heterocyclyl, wherein each of said alkyl,cycloalkyl and heterocyclyl can be unsubstituted or optionallyindependently substituted with 1–4 substituents which can be the same ordifferent, each substituent being independently selected from the groupconsisting of halo, alkyl, hydroxymethyl, hydroxyethyl, hydroxypropyl,trifluoromethyl, OR⁶, NR⁶R⁷, SR⁶, SO₂R⁶, CN, SO₂N(R6R7) and NO₂; R⁵ isH, alkyl, aryl, heteroaryl, arylalkyl, cycloalkyl, heterocyclyl, acyl orheteroarylalkyl; R⁶ is H, alkyl or aryl; R⁷ is H or alkyl; R¹⁰ is halo,alkyl, hydroxyalkyl, trifluoromethyl, OR⁶, NR⁸R⁷, SR⁶, SO₂R⁶, CN,SO₂N(R⁶R⁷) or NO₂; and n is 0 to 4, and when n is 2–4, the n moietiescan be the same or different, each being independently selected, withthe following provisos: (i) that when R² is C₁–C₄ alkyl and R⁵ is H,then R⁴ is not a C₁–C₄ alkyl; (ii) that when R² is halo, CN, formyl,nitro, alkylcarbonyl, aralkylcarbonyl, heteroaralkylcarbonyl, or-alkylene-N(R⁸R⁹), then: (a) R³ is not H, alkylthio, aralkylthio,alkylsulfinyl, aralkylsulfinyl, or —NR⁴R⁵, and (b) n is not 0; and (iii)that when R² is alkyl, cycloalkyl, alkenyl or alkynyl, then R³ is notNH(methyl), N ,N(dimethyl), NH(acetyl), N(methyl)(acetyl), H, alkyl,alkylthio, aralkylthio, alkylsulfinyl, or aralkylsulfinyl.
 2. Thepharmaceutical composition of claim 1, additionally comprising one ormore agents selected from the group consisting of cytostatic agent,cisplatin, doxorubicin, taxotere, taxol, etoposide, irinotecan,camptostar, topotecan, paclitaxel, docetaxel, epothilones, tamoxifen,5-fluorouracil, methoxtrexate, 5FU, temozolomide, cyclophosphamide, SCH66336, R115777, L778,123, BMS 214662, Iressa, Tarceva, antibodies toEGFR, Gleevec, intron, ara-C, adriamycin, cytoxan, gemcitabine, Uracilmustard, Chlormethine, Ifosfamide, Melphalan, Chlorambucil, Pipobroman,Triethylenemelamine, Triethylenethiophosphoramine, Busulfan, Carmustine,Lomustine, Streptozocin, Dacarbazine, Floxuridine, Cytarabine,6-Mercaptopurine, 6-Thioguanine, Fludarabine phosphate, Pentostatine,Vinblastine, Vincristine, Vindesine, Bleomycin, Dactinomycin,Daunorubicin, Epirubicin, Idarubicin, Mithramycin, Deoxycoformycin,Mitomycin-C, L-Asparaginase, Teniposide 17α-Ethinylestradiol,Diethylstilbestrol, Testosterone, Prednisone, Fluoxymesterone,Dromostanolone propionate, Testolactone, Megestrolacetate,Methylprednisolone, Methyltestosterone, Prednisolone, Triamcinolone,Chlorotrianisene, Hydroxyprogesterone, Aminoglutethimide, Estramustine,Medroxyprogesteroneacetate, Leuprolide, Flutamide, Toremifene,goserelin, Carboplatin, Hydroxyurea, Amsacrine, Procarbazine, Mitotane,Mitoxantrone, Levamisole, Navelbene, CPT-11, Anastrazole, Letrazole,Capecitabine, Reloxafine, Droloxafine, or Hexamethylmelamine.
 3. Apharmaceutical composition comprising one or more compounds selectedfrom:

or a pharmaceutically acceptable salt, or ester thereof.
 4. Thepharmaceutical composition of claim 3, additionally comprising one ormore agents selected from the group consisting of cytostatic agent,cisplatin, doxorubicin, taxotere, taxol, etoposide, irinotecan,camptostar, topotecan, paclitaxel, docetaxel, epothilones, tamoxifen,5-fluorouracil, methoxtrexate, 5FU, temozolomide, cyclophosphamide, SCH66336, R115777, L778,123, BMS 214662, Iressa, Tarceva, antibodies toEGFR, Gleevec, intron, ara-C, adriamycin, cytoxan, gemcitabine, Uracil ,Chlormethine, Ifosfamide, Melphalan, Chlorambucil, Pipobroman,Triethylenemelamine, Triethylenethiophosphoramine, Busulfan, Carmustine,Lomustine, Streptozocin, Dacarbazine, Floxuridine, Cytarabine,6-Mercaptopurine, 6-Thioguanine, Fludarabine phosphate, Pentostatine,Vinblastine, Vincristine, Vindesine, Bleomycin, Dactinomycin,Daunorubicin, Epirubicin, Idarubicin, Mithramycin, Deoxycoformycin,Mitomycin-C, L-Asparaginase, Teniposide 17α-Ethinylestradiol,Diethylstilbestrol, Testosterone, Prednisone, Fluoxymesterone,Dromostanolone propionate, Testolactone, Megestrolacetate,Methylprednisolone, Methyltestosterone, Prednisolone, Triamcinolone,Chlorotrianisene, Hydroxyprogesterone, Aminoglutethimide, Estramustine,Medroxyprogesteroneacetate, Leuprolide, Flutamide, Toremifene,goserelin, Carboplatin, Hydroxyurea, Amsacrine, Procarbazine, Mitotane,Mitoxantrone, Levamisole, Navelbene, CPT-11, Anastrazole, Letrazole,Capecitabine, Reloxafine, Droloxafine, or Hexamethylmelamine.